Identification, characterization, and cloning of a novel aminoacylase, L-pipecolic acid acylase from <i>Pseudomonas</i> species
-
- Hayashi Junji
- Faculty of Bioscience and Bioindustry, Tokushima University
-
- Ichiki Yoshiaki
- Department of Biotechnology, College of Life Sciences, Ritsumeikan University
-
- Kanda Akiko
- Department of Biotechnology, College of Life Sciences, Ritsumeikan University
-
- Takagi Kazuyoshi
- Department of Applied Chemistry, College of Life Sciences, Ritsumeikan University
-
- Wakayama Mamoru
- Department of Biotechnology, College of Life Sciences, Ritsumeikan University
この論文をさがす
抄録
<p>L-Pipecolic acid is utilized as a vital component of specific chemical compounds, such as immunosuppressive drugs, anticancer reagents, and anesthetic reagents. We isolated and characterized a novel L-aminoacylase, N-acetyl-L-pipecolic acid-specific aminoacylase (LpipACY), from Pseudomonas sp. AK2. The subunit molecular mass of LpipACY was 45 kDa and was assumed to be a homooctamer in solution. The enzyme exhibited high substrate specificity toward N-acetyl-L-pipecolic acid and a high activity for N-acetyl-L-pipecolic acid and N-acetyl-L-proline. This enzyme was stable at a high temperature (60°C for 10 min) and under an alkaline pH (6.0–11.5). The N-terminal and internal amino acid sequences of the purified enzyme were STTANTLILRNG and IMASGGV, respectively. These sequences are highly consistent with those of uncharacterized proteins from Pseudomonas species, such as amidohydrolase and peptidase. We also cloned and overexpressed the gene coding LpipACY in Escherichia coli. Moreover, the recombinant LpipACY exhibited properties similar to native enzyme. Our results suggest that LpipACY is a potential enzyme for the enzymatic synthesis of L-pipecolic acid. This study provides the first description of the enzymatic characterization of L-pipecolic acid specific amino acid acylase.</p>
収録刊行物
-
- The Journal of General and Applied Microbiology
-
The Journal of General and Applied Microbiology 67 (5), 186-194, 2021
公益財団法人 応用微生物学・分子細胞生物学研究奨励会